المؤلفون: Wataru Tsuchiya, Kenjiro Katsu, Kiyoshi Furukawa, Yasuo Yasui, Toshimasa Yamazaki, Seiji Kubota, Toshiyuki Komori, Amanda R. Walker, Tomomi Hisano, Rintaro Suzuki, Noritoshi Inagaki, Katsuhiro Matsui, Motoyuki Koshio

المصدر: SC10201804160188
NARO成果DBa
This is an open-access article distributed. CC BY4.0
BMC Plant Biology, Vol 17, Iss 1, Pp 1-14 (2017)
BMC Plant Biology

مصطلحات موضوعية: 0106 biological sciences, 0301 basic medicine, Recombinant protein, Population, Sequence alignment, Plant Science, Biology, 01 natural sciences, Substrate Specificity, Anthocyanins, 03 medical and health sciences, lcsh:Botany, Complementary DNA, Proanthocyanidins, Amino Acid Sequence, Asparagine, Fagopyrum esculentum, education, Peptide sequence, Phylogeny, Plant Proteins, chemistry.chemical_classification, education.field_of_study, Substrate (chemistry), biology.organism_classification, lcsh:QK1-989, Amino acid, 3D structure modelling, Alcohol Oxidoreductases, 030104 developmental biology, chemistry, Biochemistry, Flavonoid, Substrate preference, Sequence Alignment, Fagopyrum, Research Article, 010606 plant biology & botany

الوصف: Background Dihydroflavonol 4-reductase (DFR) is the key enzyme committed to anthocyanin and proanthocyanidin biosynthesis in the flavonoid biosynthetic pathway. DFR proteins can catalyse mainly the three substrates (dihydrokaempferol, dihydroquercetin, and dihydromyricetin), and show different substrate preferences. Although relationships between the substrate preference and amino acids in the region responsible for substrate specificity have been investigated in several plant species, the molecular basis of the substrate preference of DFR is not yet fully understood. Results By using degenerate primers in a PCR, we isolated two cDNA clones that encoded DFR in buckwheat (Fagopyrum esculentum). Based on sequence similarity, one cDNA clone (FeDFR1a) was identical to the FeDFR in DNA databases (DDBJ/Gen Bank/EMBL). The other cDNA clone, FeDFR2, had a similar sequence to FeDFR1a, but a different exon-intron structure. Linkage analysis in an F2 segregating population showed that the two loci were linked. Unlike common DFR proteins in other plant species, FeDFR2 contained a valine instead of the typical asparagine at the third position and an extra glycine between sites 6 and 7 in the region that determines substrate specificity, and showed less activity against dihydrokaempferol than did FeDFR1a with an asparagine at the third position. Our 3D model suggested that the third residue and its neighbouring residues contribute to substrate specificity. FeDFR1a was expressed in all organs that we investigated, whereas FeDFR2 was preferentially expressed in roots and seeds. Conclusions We isolated two buckwheat cDNA clones of DFR genes. FeDFR2 has unique structural and functional features that differ from those of previously reported DFRs in other plants. The 3D model suggested that not only the amino acid at the third position but also its neighbouring residues that are involved in the formation of the substrate-binding pocket play important roles in determining substrate preferences. The unique characteristics of FeDFR2 would provide a useful tool for future studies on the substrate specificity and organ-specific expression of DFRs. Electronic supplementary material The online version of this article (10.1186/s12870-017-1200-6) contains supplementary material, which is available to authorized users.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5d6e76e5db894307ac4bf2decac312c9Test
https://doi.org/10.1186/s12870-017-1200-6Test

المؤلفون: Shigehiro Yamada, Toshiyuki Komori, Tomoaki Kubo, Shigeru Kuwata, Patricia N. Myers, Hidemasa Imaseki

المصدر: Plant and Cell Physiology. 38:1226-1231

مصطلحات موضوعية: Physiology, Molecular Sequence Data, Aquaporin, Plant Science, Aquaporins, Ion Channels, Gene Expression Regulation, Plant, Tobacco, Gene expression, Gene family, Amino Acid Sequence, Gene, Plant Proteins, chemistry.chemical_classification, Messenger RNA, Sequence Homology, Amino Acid, biology, Water, Cell Biology, General Medicine, Membrane transport, biology.organism_classification, Amino acid, Plants, Toxic, chemistry, Biochemistry, Solanaceae

الوصف: Deduced amino acid sequences encoded by the cDNAs related to the MIP gene family from Nicotiana excelsior were characterized. Phylogenetic characterization of the products of corresponding genes named NeMip1, NeMip2, and NeMip3 strongly suggested that they are water channel proteins localized in the plasma membrane. Organ specificity of the gene expression was examined in leaves, roots, and reproductive organs. NeMip1 was expressed in roots and reproductive organs; however, it was hardly detectable in leaves. Two other genes, NeMip2 and NeMip3, were expressed in all of organs examined. mRNA accumulation from the genes was investigated in leaves under salt- and drought-stresses. The results demonstrated that mRNA accumulation from all three genes increased under salt- and drought-stresses within one day. However, they showed different accumulation patterns. In addition to their up-regulation under salt- and drought-stresses, daily changes in NeMip2 and NeMip3 mRNA accumulation was observed under unstressed conditions in leaves.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::66513e33eac4e76b520d0b03a25cab8aTest
https://doi.org/10.1093/oxfordjournals.pcp.a029109Test

المؤلفون: Toshiyuki Komori, Shigehiro Yamada, Hidemasa Imaseki, Tomoaki Kubo, Shigeru Kuwata, Akiko Hashimoto

المصدر: Plant science : an international journal of experimental plant biology. 154(1)

مصطلحات موضوعية: Messenger RNA, biology, fungi, Aldolase A, food and beverages, Fructose-bisphosphate aldolase, Plant Science, General Medicine, biology.organism_classification, Isozyme, Biochemistry, Gene expression, Genetics, biology.protein, Agronomy and Crop Science, Gene, Southern blot, Nicotiana

الوصف: Two homologous genes of plastidic fructose-1,6-bisphosphate aldolase (AldP) isozymes were isolated from green leaves of a salt stress-tolerant Nicotiana species, Nicotiana paniculata, by differential screening. The products of the corresponding genes, NpAldP1 and NpAldP2, were 91% identical to each other and 70-85% identical to the other known plant plastidic aldolases. Although these two genes showed similar organ-specific expression and daily cycles, their responses to salt stress differed: mRNA accumulation of NpAldP2 increased, but that of NpAldP1 slightly decreased. The mRNA accumulations of their counterparts of two other Nicotiana species, NeAldP1 and NeAldP2 (Nicotiana excelsior), and NaAldP1 and NaAldP2 (Nicotiana arentsii) were studied under the same stress condition. N. arentsii conserved accumulation profiles similar to N. paniculata, but N. excelsior did not. In N. excelsior, accumulation of NeAldP1 decreased to 50% of the control after stress and gradually recovered thereafter, whereas accumulation of NeAldP2 temporarily decreased and reached 250% of the control by the third day of stress. Southern blot analysis indicated that NpAldP1, NpAldP2, NaAldP1, and NaAldP2 include one or two closely related genes and NeAldP1 and NeAldP2 several.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ea94101b8ec224c6d42926fac0893ed9Test
https://pubmed.ncbi.nlm.nih.gov/10725559Test